Accumulation of mitochondrial DNA mutations in human immunodeficiency virus-infected patients treated with nucleoside-analogue reverse-transcriptase inhibitors
article
Nucleoside reverse-transcriptase inhibitor (NRTI) therapy for human immunodeficiency virus (HIV) infection has been associated with mitochondrial DNA (mtDNA) polymerase-γ inhibition and subsequent mtDNA depletion. Effects on mtDNA mutation, although suggested by critical involvement of polymerase-γ in DNA-repair reactions, are unknown. In the present study, we assessed the nature and frequency of mitochondrial genome sequence differences in peripheral-blood samples taken prior to NRTI therapy and after 6-77 mo of treatment in 16 NRTI-treated patients. Samples from 10 HIV-infected, treatment-naive control individuals were taken at similar time intervals. Single-stranded conformation polymorphism (SSCP) and DNA-sequencing analysis techniques were used to detect mitochondrial genome sequence variants between paired longitudinal samples, and heteroplasmic populations were quantified after cloning and repeat SSCP/sequencing. Of 16 individuals treated with NRTIs, 5 exhibited altered SSCP profiles associated with the development of novel heteroplasmic DNA sequence changes, whereas no SSCP pattern change within these regions was observed in the control individuals. Heteroplasmic sequence changes were distributed across four regions of the genome: the noncoding region to 12S ribosomal RNA, reduced-nicotinamide-adenine-dinucleotide dehydrogenase 1, and cytochrome oxidase subunits I and III. Of the total of 26 patients who were examined in the present study, 4 of 5 patients with detectable mtDNA sequence changes since commencement of therapy developed evidence of peripheral fat wasting (lipoatrophy) between sample intervals (P = .031). One patient, without detectable sequence changes on NRTI therapy, also developed lipoatrophy. Levels of mtDNA copies/cell in blood samples were determined by quantitative PCR for 11 of the 16 NRTI-exposed patients; 7 of these 11 patients showed reduced levels of mtDNA in blood after therapy, including all 3 patients tested with evidence of mtDNA sequence changes on therapy. These data indicate that NRTI therapy provides conditions permissive for the development of peripheral-blood mtDNA mutations in vivo. Molecular Sequence Numbers: GENBANK: NC_001807; Chemicals/CAS: cytochrome c oxidase, 72841-18-0, 9001-16-5; didanosine, 69655-05-6; indinavir, 150378-17-9, 157810-81-6, 180683-37-8; lamivudine, 134678-17-4, 134680-32-3; nevirapine, 129618-40-2; reduced nicotinamide adenine dinucleotide dehydrogenase, 9027-14-9, 9032-21-7, 9079-67-8; stavudine, 3056-17-5; zidovudine, 30516-87-1; DNA polymerase gamma, EC 2.7.7.-; DNA Primers; DNA, Mitochondrial; DNA, Ribosomal; DNA-Directed DNA Polymerase, EC 2.7.7.7; Lamivudine, 134678-17-4; Reverse Transcriptase Inhibitors; RNA, Ribosomal; RNA, ribosomal, 12S; Stavudine, 3056-17-5
Topics
cytochrome c oxidasedidanosineindinavirmitochondrial DNAnevirapinenucleoside analogreduced nicotinamide adenine dinucleotide dehydrogenaseribosome RNARNA directed DNA polymerase inhibitorzidovudineblood analysisclinical articleclinical trialcontrolled clinical trialcontrolled studyDNA sequenceenzyme subunitfemalegene frequencygene mutationgene sequencegene structureheteroplasmyHuman immunodeficiency virusHuman immunodeficiency virus infectionlipoatrophymalemitochondrial geneticsmolecular cloningnonhumannucleotide sequencepolymerase chain reactionrandomized controlled trialsingle strand conformation polymorphismBase SequenceDNA Mutational AnalysisDNA PrimersDNA, MitochondrialDNA, RibosomalDNA-Directed DNA PolymeraseHIV InfectionsHumansLamivudineMolecular Sequence DataMutagenesisPolymorphism, Single-Stranded ConformationalReverse Transcriptase InhibitorsRNA, RibosomalStavudineTime FactorsHuman immunodeficiency virusRNA viruses
TNO Identifier
236990
ISSN
00029297
Source
American Journal of Human Genetics, 72(3), pp. 549-560.
Pages
549-560
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